Patent US3991759A successfully refreshed from Google Patents.
EP0244221A1
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Hybrid receptors, nucleic acid encoding them, their preparation, and their use in determination of ligands and their antagonists and agonists
Assignee: Genentech Inc
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Filing: 1987-04-29
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Granted: —
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Expires: 2007-04-29
Abstract
Abstract Hybrid receptors, produced by recombinant DNA technology, comprise (a) the ligand binding domain of a predetermined receptor and (b) a heterologous reporter polypeptide. The hybrid receptors are useful for convenient and large scale assay of biologically active ligands or their antagonists or agonists. (a) may be the extracellular domain of the receptor, or a cytoplasmic domain of a receptor or oncogene. (b) may be an enzyme. A transmembrane domain may be interposed between (a) and (b).
Inventors
- 👤 Thomas Joseph Dull
- 👤 Heimo Riedel
- 👤 Axel Ullrich
Bibliographic Details
Application Number
—
Jurisdiction
EP
Data Source
google_patents
Last Synchronized
2026-10-23 22:54
CPC Classifications
C; C07; C07K; C07K14/00; C07K14/435; C07K14/705; C07K14/72; C07K14/71; G; G01; G01N; G01N33/00; G01N33/48; G01N33/50; G01N33/53; G01N33/531; G01N33/532; G01N33/566; G01N33/68; C07K2319/00; C07K2319/01; C07K2319/033; C07K2319/32; C07K2319/90; G01N2500/00; Y; Y10; Y10S; Y10S435/00; Y10S435/968; Y10S530/00; Y10S530/806; Y10S530/808
Claims Analysis
Total Claims: 53
Claim 1
Independent Claim
19 words
A hybrid receptor comprising (a) the ligand binding domain of a predetermined receptor and (b) a heterologous reporter polypeptide.
Claim 2
Dependent Claim
19 words
A hybrid receptor comprising (a) the ligand binding domain of a predetermined receptor and (b) a heterologous reporter polypeptide.
Claim 3
Dependent Claim
21 words
The hybrid receptor of claim 1 wherein the ligand binding domain is comprised by the extracellular domain of the predetermined receptor.
Claim 4
Dependent Claim
19 words
The hybrid receptor of claim 1 wherein the reporter polypeptide is a cytoplasmic domain of a receptor or oncogene.
Claim 5
Dependent Claim
13 words
The hybrid receptor of claim 1 wherein the reporter polypeptide is an enzyme.
Claim 6
Dependent Claim
23 words
The hybrid receptor of claim 4 wherein the enzyme is not sterically inhibited by the binding of a ligand to the hybrid receptor.
Claim 7
Dependent Claim
12 words
The hybrid receptor of claim 4 wherein the enzyme is a phosphorylkinase.
Claim 8
Dependent Claim
21 words
The hybrid receptor of claim 1 having a transmembrane domain interposed between the ligand binding domain and the heterologous reporter polypeptide.
Claim 9
Dependent Claim
22 words
Nucleic acid encoding a hybrid receptor comprising (a) the ligand binding domain of a predetermined receptor and (b) a heterologous reporter polypeptide.
Claim 10
Dependent Claim
22 words
Nucleic acid encoding a hybrid receptor comprising (a) the ligand binding domain of a predetermined receptor and (b) a heterologous reporter polypeptide.
Claim 11
Dependent Claim
11 words
The nucleic acid of claim 8 further comprising a replicable vector.
Claim 12
Dependent Claim
10 words
The vector of claim 9 further comprising a host cell.
Claim 13
Independent Claim
69 words
A method for making a hybrid receptor which receptor comprises (1) the ligand binding domain of a predetermined receptor and (2) a heterologous reporter polypeptide, said method comprising: (a) transforming a host cell with a vector containing nucleic acid encoding the hybrid receptor operably linked to a promoter for controlling the transcription of the hybrid receptor; and (b) culturing the host cell under conditions for expressing the hybrid receptor.
Claim 14
Independent Claim
69 words
A method for making a hybrid receptor which receptor comprises (1) the ligand binding domain of a predetermined receptor and (2) a heterologous reporter polypeptide, said method comprising: (a) transforming a host cell with a vector containing nucleic acid encoding the hybrid receptor operably linked to a promoter for controlling the transcription of the hybrid receptor; and (b) culturing the host cell under conditions for expressing the hybrid receptor.
Claim 15
Dependent Claim
19 words
The method of claim 11 wherein the hybrid receptor is recovered from the culture medium of the host cell.
Claim 16
Dependent Claim
19 words
The method of claim 11 wherein the hybrid receptor is recovered from the cell membrane of the host cell.
Claim 17
Independent Claim
82 words
A method for assaying a biologically active ligand or an antagonist or agonist for said ligand, comprising: (a) providing a hybrid receptor which comprises (1) a binding domain for the ligand, antagonist or agonist and (2) a heterologous reporter polypeptide; (b) incubating the receptor with a test sample suspected to contain the ligand, antagonist or agonist; (c) detecting a change in the reporter polypeptide; and (d) correlating said change with the presence of the ligand, antagonist or agonist in the test sample.
Claim 18
Independent Claim
82 words
A method for assaying a biologically active ligand or an antagonist or agonist for said ligand, comprising: (a) providing a hybrid receptor which comprises (1) a binding domain for the ligand, antagonist or agonist and (2) a heterologous reporter polypeptide; (b) incubating the receptor with a test sample suspected to contain the ligand, antagonist or agonist; (c) detecting a change in the reporter polypeptide; and (d) correlating said change with the presence of the ligand, antagonist or agonist in the test sample.
Claim 19
Dependent Claim
25 words
The method of claim 14 wherein the ligand is a polypeptide and the ligand binding domain is not the antigen binding site of an immunoglobulin.
Claim 20
Dependent Claim
22 words
The method of claim 14 wherein the change in the reporter polypeptide is a modification of the enzymatic activity of the polypeptide.
Claim 21
Dependent Claim
18 words
The method of claim 16 wherein the, change in the reporter polypeptide is autophosphorylation of the reporter polypeptide.
Claim 22
Dependent Claim
33 words
The method of claim 14 wherein the test sample is suspected to contain an antagonist and the receptor is incubated with the test sample and a predetermined activity of ligand or ligand agonist.
Claim 23
Dependent Claim
20 words
19: The method of claim 14 wherein the change in the reporter polypeptide is a change in an immune epitope.
Claim 24
Dependent Claim
37 words
The method of claim 19 wherein the change in immune epitope is detected by incubating the receptor with an antibody capable of binding to the reporter polypeptide and determining the amount of bound or residual unbound polypeptide.
Claim 25
Dependent Claim
56 words
The method of claim 14 wherein the reporter polypeptide further comprises a stable free radical, fluorescent or chemiluminescent group and the change in the reporter polypeptide is detected by measuring a change in the rotational moment of the stable free radical or a change in the intensity, wavelength or polarization of the fluorescent or chemiluminescent group.
Claim 26
Dependent Claim
17 words
The method of claim 14 wherein the reporter polypeptide is capable of binding to a G protein.
Claim 27
Dependent Claim
26 words
A process which comprises the preparation of a hybrid receptor comprising (a) the ligand binding domain of a predetermined receptor and (b) a heterologous reporter polypeptide.
Claim 28
Dependent Claim
26 words
A process which comprises the preparation of a hybrid receptor comprising (a) the ligand binding domain of a predetermined receptor and (b) a heterologous reporter polypeptide.
Claim 29
Dependent Claim
20 words
The process of claim 1 wherein the ligand binding domain is comprised by the extracellular domain of the predetermined receptor.
Claim 30
Dependent Claim
18 words
The process of claim 1 wherein the reporter polypeptide is a cytoplasmic domain of a receptor or oncogene.
Claim 31
Dependent Claim
13 words
The process of ciaim 1 wherein the reporter polypeptide is an enzyme. I
Claim 32
Dependent Claim
13 words
The process of ciaim 1 wherein the reporter polypeptide is an enzyme. I
Claim 33
Dependent Claim
22 words
The process of claim 4 wherein the enzyme is not sterically inhibited by the binding of a ligand to the hybrid receptor.
Claim 34
Dependent Claim
12 words
The process of claim 4 wherein the enzyme ' is a phosphorylkinase.
Claim 35
Dependent Claim
24 words
The process of claim 1 wherein the hybrid receptor has a transmembrane domain interposed between the ligand binding domain and the heterologous reporter polypeptide.
Claim 36
Dependent Claim
29 words
A process which comprises the preparation of nucleic acid encoding a hybrid receptor comprising (a) the ligand binding domain of a predetermined receptor and (b) a heterologous reporter polypeptide.
Claim 37
Dependent Claim
29 words
A process which comprises the preparation of nucleic acid encoding a hybrid receptor comprising (a) the ligand binding domain of a predetermined receptor and (b) a heterologous reporter polypeptide.
Claim 38
Dependent Claim
14 words
The process of claim 8 wherein the nucleic acid further comprises a replicable vector.
Claim 39
Dependent Claim
15 words
The process of claim 9 which further comprises transforming a host cell with the vector.
Claim 40
Independent Claim
69 words
A method for making a hybrid receptor which receptor comprises (1) the ligand binding domain of a predetermined receptor and (2) a heterologous reporter polypeptide, said method comprising: (a) transforming a host cell with a vector containing nucleic acid encoding the hybrid receptor operably linked to a promoter for controlling the transcription of the hybrid receptor; and (b) culturing the host cell under conditions for expressing the hybrid receptor.
Claim 41
Independent Claim
69 words
A method for making a hybrid receptor which receptor comprises (1) the ligand binding domain of a predetermined receptor and (2) a heterologous reporter polypeptide, said method comprising: (a) transforming a host cell with a vector containing nucleic acid encoding the hybrid receptor operably linked to a promoter for controlling the transcription of the hybrid receptor; and (b) culturing the host cell under conditions for expressing the hybrid receptor.
Claim 42
Dependent Claim
19 words
The method of claim 11 wherein the hybrid receptor is recovered from the culture medium of the host cell.
Claim 43
Dependent Claim
19 words
The method of claim 11 wherein the hybrid receptor is recovered from the cell membrane of the host cell.
Claim 44
Independent Claim
82 words
A method for assaying a biologically active ligand or an antagonist or agonist for said ligand, comprising: (a) providing a hybrid receptor which comprises (1) a binding domain for the ligand, antagonist or agonist and (2) a heterologous reporter polypeptide; (b) incubating the receptor with a test sample suspected to contain the ligand, antagonist or agonist; (c) detecting a change in the reporter polypeptide; and (d) correlating said change with the presence of the ligand, antagonist or agonist in the test sample.
Claim 45
Independent Claim
82 words
A method for assaying a biologically active ligand or an antagonist or agonist for said ligand, comprising: (a) providing a hybrid receptor which comprises (1) a binding domain for the ligand, antagonist or agonist and (2) a heterologous reporter polypeptide; (b) incubating the receptor with a test sample suspected to contain the ligand, antagonist or agonist; (c) detecting a change in the reporter polypeptide; and (d) correlating said change with the presence of the ligand, antagonist or agonist in the test sample.
Claim 46
Dependent Claim
25 words
The method of claim 14 wherein the ligand is a polypeptide and the ligand binding domain is not the antigen binding site of an immunoglobulin.
Claim 47
Dependent Claim
22 words
The method of claim 14 wherein the change in the reporter polypeptide is a modification of the enzymatic activity of the polypeptide.
Claim 48
Dependent Claim
18 words
The method of claim 16 wherein the change in the reporter polypeptide is autophosphorylation of the reporter polypeptide.
Claim 49
Dependent Claim
33 words
The method of claim 14 wherein the test sample is suspected to contain an antagonist and the receptor is incubated with the test sample and a predetermined activity of ligand or ligand agonist.
Claim 50
Dependent Claim
19 words
The method of claim 14 wherein the change in the reporter polypeptide is a change in an immune epitope.
Claim 51
Dependent Claim
37 words
The method of claim 19 wherein the change in immune epitope is detected by incubating the receptor with an antibody capable of binding to the reporter polypeptide and determining the amount of bound or residual unbound polypeptide.
Claim 52
Dependent Claim
56 words
The method of claim 14 wherein the reporter polypeptide further comprises a stable free radical, fluorescent or chemiluminescent group and the change in the reporter polypeptide is detected by measuring a change in the rotational moment of the stable free radical or a change in the intensity, wavelength or polarization of the fluorescent or chemiluminescent group.
Claim 53
Dependent Claim
17 words
The method of claim 14 wherein the reporter polypeptide is capable of binding to a G protein.
References Cited (Backward Citations)
DE3100061A1
Cited
US4504587A
Cited
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